Identification of triacylglycerols by high-performance liquid chromatography-gas-liquid chromatography and liquid chromatography-mass spectrometry.

Triacylglycerols from rat adipose tissue were chromatographed by high-performance liquid chromatography (HPLC), with a gradient of propan-2-ol in acetonitrile as the mobile phase. Fractions of the material eluting from the column were collected and analysed by automated gas - liquid chromatography of the fatty acid methyl esters obtained after transmethylation. Triacylglycerols were identified by using a combination of their fatty acid content and elution time from the HPLC column. Fractions corresponding to whole peaks or groups of peaks were also collected and re-chromatographed on a liquid chromatography - mass spectrometry system equipped with a belt interface. For most triacylglycerols, good agreement was obtained between the two methods, although mass spectrometric identification of the early eluting peaks was complicated by poor resolution of the triacylglycerols on the HPLC system.

Analysis of triacylglycerols by combined high-performance liquid chromatography and gas-liquid chromatography.

A method for the analysis of the triacylglycerols from animal and human adipose tissue has been developed by combining high-performance liquid chromatography and gas-liquid chromatography (HPLC) and (GLC). Following isolation by thin-layer chromatography, the triacylglycerols are chromatographed by HPLC on a 5-micron Spherisorb ODS column with a gradient of 45 to 60% 2-propanol in acetonitrile as the mobile phase. Fractions eluted from the column are collected at intervals ranging from 0.2 to 1.0 min, and the triacylglycerols present are then trans-esterified prior to analysis of the resulting fatty acid methyl esters by an automated packed column GLC system. Triacylglycerols eluted from the column may be identified by a combination of their elution volume in HPLC and their fatty acid content. From the GLC data, profiles can be constructed for the distribution of several fatty acids amongst the triacylglycerols present in the adipose tissue sample. The method can be used with solvents such as acetonitrile, 2-propanol and acetone in the mobile phase, and there is no baseline drift during gradient elution. The adipose tissue triacylglycerols of normal and streptozotocin induced diabetic rats have been compared. No differences in the distribution of the fatty acids in the two groups of rats were observed under the conditions of our study.